1. Field of the Invention
This invention relates to an herbal composition for the treatment and remedy of bronchial respiratory difficulties. More particularly this invention describes the process of separation, physicochemical characterization and biological response evaluation of active components obtained from extracts of any plant parts including leaves, barks, roots and seeds of plants M. koenigii and P. betle in order to establish their role on the treatment and remedy of bronchial respiratory troubles.
2. Background and Prior Art References
Respiratory problem consists of mild to extremely severe trouble of breathing along with the other discomforts such as wheezing, coughing, chest tightness and the like. In spite of precautionary measure, public awareness campaign and monitoring system, population having respiratory trouble is on the rise all over the world. This is true in western advanced countries, especially among the children. Using M. koenigii leaf preparation, the relief and possible cure of asthma has already been demonstrated by the applicants in their PCT Patent application No: PCT/IN/00102 filed on dated Oct. 16, 2000. Two-prong strategy is adapted in the present invention.
The respiratory disease is the result of pathophysiological symptoms arising out of aberration of immune system. The immediate symptom includes bronchial constriction, inflammation of respiratory tract and closing of air way by mucus secretion. The symptomatic drugs provide relief by temporary relaxation of the distressed symptoms.
The root cause for respiratory problems is not well addressed by the developers of symptomatic drugs. With the advent of current knowledge, it is now well accepted that leukotrienes are found to be the main player in developing symptoms of respiratory problems.
The major symptoms of respiratory problem can be divided into early and late responses. The early response occurs within minutes of allergen exposure and involves primarily mediators such as histamine, leukotrienes and prostaglandin D2. The effects of these mediators result in bronchocontriction and accumulation of mucus. The late response occurs hours later and involves additional mediators including IL-4, IL-5, IL-6, and TNF-alpha, eosinophils chemotactic factor (ECF) and platelet aggregation factor (PAF). The overall effect of these mediators is to recruit inflammatory cells including eosinophils and neutrophils. These cells are capable of causing significant tissue injury by releasing toxic enzymes. These events lead to occlusion of bronchial lumen with mucus protein, and cellular debris, thus thickening of basement membrane, fluid build up and hypertrophy of the bronchial smooth muscle. A mucus plug often forms and adheres to the bronchial wall. The mucus plug contains clusters of detached epithelial cells fragments, eosinophils, some neutrophils and spirals of bronchial tissue known as Curschmann""s spiral (Immunology, J. Kuby; W. H. Freeman and Co., New York; 3rd edition 1997). In view of the current mechanisms regarding manifestation of bronchial asthma/bronchial respiratory problems, modern strategy for drug development stressed the following approaches:
These include i) inhibition of leukotriene synthesis via blocking the 5-lipoxygenase-enzyme activity (Leqqis R A et al New England J. Med. 323:, 645,1990). The formation of leukotrienes originates from the oxidation of arachidonic acid, hence inhibition of this reaction leads to the inhibition of leukotriene synthesis. Besides, leukotriene receptors antagonists have also been introduced as anti leukotriene therapy for asthma/respiratory problems (Tien F. C., Medical J. Aust. 171: 378,1999). Currently licensed drug zelutin, based on inhibition of arachidonic acid oxidation has already been introduced exclusively in the US market. However, its use is limited by hepatotoxicity. Children below 14 years are not recommended for this drug. Moreover, patients taking other drugs need to be surveyed when taking zelutin as anti-asthma drug. More over patients taking other drugs, need to be surveyed when taking zelutin, as anti-asthma drug. ii) the neutralization of IgE either by anti-IgE antibody (humanized) or by blocking the high affinity IgE receptor, Fcxcex5R-I (Heusser C, Jardiu P. Current Oppinio Immunol., 9: 805,1999). Since suppression of Th2 cytokines leads to decrease in IgE production, additional approach is based on the inhibition of these Th2 cytokine synthesis and enhancement of Th1 cytokine formation (Chung K. F.; Barens P. J. Thorax 54: 825, 1999).
In contrast, M. koenigii based anti asthmatic preparation has been tried on children as young as 7 years old and octogenarian as old as 80 years and above without any adverse reaction.
The fractions of M. koenigii extracts obtained from all plant parts including leaves, barks, roots and seeds are therefore examined to identify the active components. These are based on their potential to inhibit the production of leukotrienes. Additionally, the fractions are also examined for shifting of Th2 response towards to Th1 type. Th1 and Th2 response are measured by xcex3-interferon (Th1) and IL-4 (Th2) production respectively. Some M. koenigii fractions showed inhibition of 5-lipoxygenase mediated arachidonic acid oxidation signifying blockage of leukotriene synthesis and remarkable increase in xcex3-interferon production which in turn would suppress IL-4 production. On the other hand, most of the P. betle leaf extract-fractions resulted in facilitating of the shift from Th2 type response to Th1 type. Thus, a blend of selected extract components from these two plants M. koenigii and P. betle predominantly inhibited leukotriene synthesis and shifted Th2 response towards Th1 type and therefore is proposed as a unique synergistic composition for treatment, relief and remedy of bronchial respiratory problems Thus the two pronged strategy is the major objective of this new composition and can be considered as the best modality of treatment for patients bronchial respiratory problems.
The main object of the invention is to provide the bioactive fractions, which are obtained from the plants parts of Murraya koenigii and Piper betle. 
Another object of the invention is to provide a new composition comprising combination of active components derived from Murraya koenigii and Piper betle plant parts for treating respiratory problems.
Another object of the present invention is to provide a process for the isolation of active components from the plant parts of M. koenigii and P. betle useful for relief, treatments and remedy of respiratory problems.
Yet another objective of the present invention is to provide a simplified method of isolation of active components from all plant parts of M. koenigii and P. betle possessing biological activities relevant to treatment, relief and remedy of respiratory problems.
Yet another objective of the present invention is to provide a simplified fast and inexpensive process for the preparation of a composition possessing biological activities relevant to treatments, relief and remedy of respiratory problems.
Yet another objective of the present invention is to provide a herbal composition preparation, comprising of active factors and components derived from all plant parts of M. koenigii and P. betle, wherein the said factors and components being highly compatible for human consumption and the treatments, relief and remedy of respiratory problems.
Yet another objective of the present invention is to examine each of the active components individually or in combination from all plant parts M. koenigii and P. betle leaves having inhibitory activity of 5-lipoxygenase mediated Arachidonic acid oxidation and promoting the shift of Th2 type response towards Th1 type.
Yet another objective of the present invention is to assay 5-lipoxygenase mediated Arachidonic acid oxidation in an ex vivo whole human blood system in the presence of M. koenigii and P. betle components individually and in combination.
Yet another objective of the present invention is to assay the 5-lipoxygenase mediated Arachidonic acid oxidation in ex vivo systems with enriched human polymorphonuclear neutrophils (PMN) in the presence of M. koenigii and P. betle components individually and in combination.
Yet another objective of the present invention is to detect the production of intracellular cytokines by flowcytometry.
Yet another objective of the present invention is to provide intracellular IFN-gama and IL-4 profile as known markers for Th1 and Th2 type response respectively in ex vivo human whole blood with the components from all plant parts of M. koenigii and P. betle individually and in combination.
Accordingly, the present invention provides an herbal composition useful for the treatment and remedy of bronchial respiratory difficulties. Another aspect of the invention, particularly describes a process of separation, physicochemical characterization and biological response evaluation of active components obtained from the extracts of any plant parts including leaves, barks, roots and seeds of plants M. koenigii and P. betle in order to establish their role on the treatment and remedy of bronchial respiratory troubles.